The antibiotic novobiocin binds and activates the ATPase that powers lipopolysaccharide transport
Abstract
Novobiocin is an orally active antibiotic that inhibits DNA gyrase by binding the ATP-binding site in the ATPase subunit. Although effective against Gram-positive pathogens, novobiocin has limited activity against Gram-negative organisms due to the presence of the lipopolysaccharide-containing outer membrane, which acts as a permeability barrier. Using a novobiocin-sensitive Escherichia coli strain with a leaky outer membrane, we identified a mutant with increased resistance to novobiocin. Unexpectedly, the mutation that increases novobiocin resistance was not found to alter gyrase, but the ATPase that powers lipopolysaccharide (LPS) transport. Co-crystal structures, biochemical, and genetic evidence show novobiocin directly binds this ATPase. Novobiocin does not bind the ATP binding site but rather the interface between the ATPase subunits and the transmembrane subunits of the LPS transporter. This interaction increases the activity of the LPS transporter, which in turn alters the permeability of the outer membrane. We propose that novobiocin will be a useful tool for understanding how ATP hydrolysis is coupled to LPS transport.
Additional Information
© 2017 American Chemical Society. Received: July 24, 2017; Published: November 14, 2017. This research was supported by the NIH (R01 GM100951 to N.R.; R01 GM066174, U19 AI109764, and R01 AI081059 to D.K.), NSF GRFP (DGE-1144152 to M.D.M.), and the Blavatnik Biomedical Accelerator at Harvard University. We wish to thank the Dana Farber Cancer Institute/Harvard Medical School joint NMR Core. This work used NE-CAT beamlines (GM103403), a Pilatus detector (RR029205), and an Eiger detector (OD021527) at the APS (DE-AC02-06CH11357). The authors declare no competing financial interest.Attached Files
Supplemental Material - ja7b07736_si_001.pdf
Supplemental Material - ja7b07736_si_002.cif
Supplemental Material - ja7b07736_si_003.cif
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Additional details
- Eprint ID
- 83221
- DOI
- 10.1021/jacs.7b07736
- Resolver ID
- CaltechAUTHORS:20171115-102801445
- R01 GM100951
- NIH
- R01 GM066174
- NIH
- U19 AI109764
- NIH
- R01 AI081059
- NIH
- DGE-1144152
- NSF Graduate Research Fellowship
- Harvard University
- Dana Farber Cancer Institute
- GM103403
- NIH
- RR029205
- NIH
- OD021527
- NIH
- DE-AC02-06CH11357
- Department of Energy (DOE)
- Created
-
2017-11-15Created from EPrint's datestamp field
- Updated
-
2021-11-15Created from EPrint's last_modified field